Structural Determination of Apolipoprotein A-I/preb-HDL Particles
EMSL Project ID
10593
Abstract
Apolipoprotein A-I (apoAI) is a 243-residue exchangeable apolipoprotein that plays a key role in the high-density lipoprotein (HDL) pathway, which clears the "bad cholesterol" from human blood stream. ApoAI is the most abundant protein component in HDL (~70%). Depending on the extent of lipidation, apoAI displays a dramatic conformational plasticity and may adopt one of four distinct conformations: (1). Lipid-free apoAI, (2). ApoAI/prebHDL, (3). ApoAI/DIscHDL, (4). ApoAI/sphericalHDL. Importantly, each conformation mediates one specific biological function, which regulates HDL formation, maturation and transportation. We plan to solve one of the apoAI conformation: apoAI on the prebHDL. Previously, using our 500 MHz and PNNL's high-field NMR instruments, we collected high quality 3D-NMR data set, allowing us to achieve ~85% backbone assignment of this 243-residue protein (MW of prebetaHDL: 55 kDa). We will also collect two 4D-NMR spectral in August. In this proposal, we request more high-field NMR data so that we can finish two other 4D-NMR experiments. These are a set of 4D NMR experiments used for spectral assignment with significant spectral overlap. We anticipate that these 4D-NMR experiments will allow us to completely assign the backbone atoms. We also request 600 MHz NMR time (with a cold probe) for a 3D 15N/13C-filtered NOESY for measurement of apoAI/POPC interaction. These NMR experiments will allow us to solve the structure of apoAI/prebHDL particles.
Project Details
Project type
Capability Research
Start Date
2004-10-20
End Date
2005-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
A complete backbone spectral assignment of human apolipoprotein AI on a 38 kDa preβHDL (Lp1-AI) particle Xuefeng Ren, Yunhuang Yang, Tracey Neville, David Hoyt, Daniel Sparks, Jianjun Wang
Apolipoprotein AI tertiary structures determine stability and phospholipidbinding activity of discoidal high-density lipoprotein particles of different sizes
Bin Chen, Xuefeng Ren, Tracey Neville, Gray Jerome, David W. Hoyt, Daniel Sparks, Gang Ren, and Jianjun Wang
Received 14 November 2008; Revised 25 February 2009; Accepted 26 February 2009
DOI: 10.1002/pro.101 Published online 16 March 2009 proteinscience.org
Chen B, X Ren, T Neville, WG Jerome, DW Hoyt, DL Sparks, G Ren, and J Wang. 2009. "Apolipoprotein AI tertiary structures determine stability and phospholipid-binding activity of discoidal high-density lipoprotein particles of different sizes." Protein Science 18(5):921-935. doi:10.1002/pro.101